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Measuring Naturally Acquired Phagocytosis-Inducing Antibodies to Plasmodium falciparum Parasites by a Flow Cytometry-Based Assay
Published on: August 6, 2020
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A versatile, high through-put, bead-based phagocytosis assay for Plasmodium falciparum
Yukie M Lloyd1, Elise P Ngati1, Ali Salanti2,3
1Department of Tropical Medicine, Medical Microbiology and Pharmacology, John A. Burns School of Medicine, University of Hawaii at Manoa, Hawaii, USA.
Scientific Reports
|November 2, 2017
Summary
A new flow cytometry assay simplifies measuring phagocytosis of Plasmodium falciparum-infected cells. This method uses antigen-coupled beads for repeatable and quantifiable results, improving malaria research.
Area of Science:
- Immunology
- Parasitology
- Biotechnology
Background:
- Antibody-mediated phagocytosis is crucial for clearing Plasmodium falciparum-infected erythrocytes (IE).
- Existing phagocytosis assays for IE suffer from high variability due to reliance on patient samples or in vitro cultures.
- There is a need for a standardized, high-throughput assay for studying IE phagocytosis.
Purpose of the Study:
- To develop a simple, high-throughput, and repeatable flow cytometric assay for measuring phagocytosis of malaria-infected cells.
- To provide a semi-quantitative measure of antigen uptake in phagocytosis assays.
Main Methods:
- A novel assay utilizing THP-1 cells and fluorescent beads.
- Beads were covalently coupled with the malarial antigen VAR2CSA.
- Phagocytosis was assessed via flow cytometry, measuring both percentage and quantity of internalized beads.
Main Results:
- The developed assay is highly repeatable.
- It provides a reliable measure of the overall percentage of phagocytosis.
- The assay semi-quantitates the number of antigen-coupled beads internalized by cells.
Conclusions:
- A simple, high-throughput flow cytometric assay for IE phagocytosis has been established.
- This assay offers improved reproducibility and quantification compared to traditional methods.
- The assay facilitates more reliable research into antibody-mediated immunity against malaria.
